High resolution systems, kits, apparatus, and methods for screening microorganisms and other high throughput microbiology applications
AssigneeIsolation Bio Inc.
InventorsAlexander Hallock, Kimberly Wemmer
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Abstract
A microfabricated device defining a high density array of microwells is described for cultivating cells from a sample. The device may be incubated to grow a plurality of cells, which may be split into an analysis portion and a reserve portion. Methods are provided for screening a biological entity of interest using the microfabricated device, for example, for screening phosphate solubilizing bacteria or other bacteria producing acids.
Claims12
A method of screening for at least one biological entity of interest in a sample using a microfabricated device having a top surface defining an array of microwells, the method comprising:
- loading into at least one microwell of the array of microwells:
- (a) at least one cell from the sample;
- (b) an indicator substance;
- and (c) an amount of a nutrient;
- applying a membrane to the microfabricated device to retain the at least one cell in the at least one microwell;
- incubating the microfabricated device at predetermined conditions for a duration of time to grow a plurality of cells from the at least one cell in the at least one microwell;
- evaluating an optical property of the at least one microwell;
- and
- determining a presence or absence of at least one biological entity of interest in the at least one microwell based on the optical property;
- wherein the indicator substance comprises an inorganic phosphate salt that is insoluble in water;
- and wherein the at least one biological entity of interest comprises a microorganism that solubilizes the inorganic phosphate salt.
A method of screening for at least one biological entity of interest in a sample using a microfabricated device having a top surface defining an array of microwells, the method comprising:
- loading into at least one microwell of the array of microwells:
- (a) at least one cell from the sample;
- (b) an indicator substance;
- and (c) an amount of a nutrient;
- applying a membrane to the microfabricated device to retain the at least one cell in the at least one microwell;
- incubating the microfabricated device at predetermined conditions for a duration of time to grow a plurality of cells from the at least one cell in the at least one microwell;
- and
- determining a presence or absence of at least one biological entity of interest in the at least one microwell based on a change of the indicator substance after incubation,
- wherein the indicator substance comprises an inorganic phosphate salt that is insoluble in water, and wherein the at least one biological entity of interest comprises a microorganism that solubilizes the inorganic phosphate salt.
A method of screening for at least one biological entity of interest in a sample using a microfabricated device having a top surface defining an array of microwells, wherein each of the microwells has a bottom, the method comprising:
- depositing an indicator substance into each of a plurality of microwells of the array of microwells;
- loading the sample onto the microfabricated device such that at least one microwell of the plurality of microwells receives at least one cell from the sample and an amount of a nutrient;
- applying a membrane to the microfabricated device to retain the received at least one cell and the nutrient in each of the plurality of microwells;
- incubating the microfabricated device at predetermined conditions for a duration of time;
- and
- determining a presence or absence of at least one biological entity of interest in the at least one microwell of the plurality of microwells based on a measurement of an optical property of the at least one microwell,
- wherein the indicator substance comprises an inorganic phosphate salt that is insoluble in water, and wherein the at least one biological entity of interest comprises a microorganism that solubilizes the inorganic phosphate salt.
Dependencies are inferred from the claim text.